Mitotic index maximization with no effect on radiation-induced dicentric chromosome frequency.

IF 2.1 4区 医学 Q2 BIOLOGY International Journal of Radiation Biology Pub Date : 2023-01-01 DOI:10.1080/09553002.2023.2142981
Kai Takebayashi, Keito Echizenya, Yuki Kameya, Daichi Nakajima, Ryo Nakayama, Yohei Fujishima, Valerie Swee Ting Goh, Yu Abe, Kosuke Kasai, Donovan A Anderson, William F Blakely, Tomisato Miura
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引用次数: 2

Abstract

Purpose: The dicentric chromosome (Dic) assay, which is the gold standard for biological dose assessment in radiation emergency medicine, requires an analysis of at least 500 lymphocyte metaphases or 100 Dic aberrations. Therefore, peripheral blood culture conditions able to obtain a high frequency of metaphases for efficient dose evaluation should be optimized. However, the type of blood cultures [i.e. whole blood (WB) or isolated peripheral blood mononuclear cell (PBMC)-culture] and blood volume differ between biodosimetry laboratories. The purpose of this study is to investigate the blood volume at which a high mitotic index (MI) is obtained in peripheral WB-culture and isolated PBMC-culture, and to examine the possible effect of blood volume on radiation-induced Dic frequency.

Materials and methods: Peripheral blood was collected from three healthy donors with their informed consent. The complete and differential blood counts were performed using an automated hematology analyzer. After blood count, peripheral blood was irradiated with 0 or 2 Gy X-ray. Blood was cultured with phytohemagglutinin (180 μg/ml) and demecolcine (0.05 μg/ml) for 48 h. The MI and Dic frequency were analyzed in 5, 10, 15, 20, 25, and 30% WB-cultures and 0.6, 1.2, 1.8, 2.4, 3.0, 3.6, and 4.2 ml WB-equivalent PBMC-cultures.

Results: In WB-culture, MI showed the highest value (∼22%) in 5-15% WB-culture and then gradually decreased to ∼9% with 30% WB-culture. MI peaked at 36 and 31% in 1.8 and 2.4 ml-WB equivalent volumes for PMBC-cultures, respectively. MI progressively decreased as the amount of PBMCs increased. Although individual differences were observed in the MI values among the three subjects, all the subjects showed the same tendency and higher MI was seen in PBMC than WB-cultures. However, these factors had no significant impact on the yield of Dics. In all culture conditions, the estimated dose calculated based on the Dic frequency was equivalent to the absorbed dose of ex vivo X-ray-irradiated blood.

Conclusion: While MI was affected by the blood culture type and the volume of cultured blood, Dic yield did not differ significantly between these conditions. These results could be used by relevant laboratories to optimize MI in certain circumstances.

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有丝分裂指数最大化,对辐射诱导的双中心染色体频率无影响。
目的:双中心染色体(Dic)测定是放射急救医学中生物剂量评估的金标准,需要分析至少500个淋巴细胞中期或100个Dic畸变。因此,应优化能够获得高中期频率的外周血培养条件,以进行有效的剂量评估。然而,血液培养物的类型[即全血(WB)或分离外周血单个核细胞(PBMC)培养物]和血容量在生物剂量测定实验室之间有所不同。本研究的目的是探讨外周血wb培养和离体pbmc培养中获得高有丝分裂指数(MI)的血容量,并探讨血容量对辐射诱导的Dic频率的可能影响。材料与方法:采集3名健康献血者经知情同意的外周血。全自动血液学分析仪进行全血细胞计数和差异血细胞计数。血球计数后,用0或2 Gy x线照射外周血。用植物血凝素(180 μg/ml)和去胆碱(0.05 μg/ml)培养48 h。分析5、10、15、20、25和30% wb -当量培养以及0.6、1.2、1.8、2.4、3.0、3.6和4.2 ml wb -当量pbmc培养的心肌梗死和Dic频率。结果:在wb培养中,MI在5-15% wb培养中最高(约22%),在30% wb培养中逐渐下降至约9%。pmbc培养物的等效体积分别为1.8 ml-WB和2.4 ml-WB时,MI峰值分别为36%和31%。心肌梗死随着pbmc数量的增加而逐渐降低。虽然三个被试之间的MI值存在个体差异,但所有被试都表现出相同的趋势,PBMC的MI值高于wb培养。然而,这些因素对Dics的产率没有显著影响。在所有培养条件下,基于Dic频率计算的估计剂量与体外x射线照射血液的吸收剂量相当。结论:心肌梗死受血培养类型和培养血容量的影响,但心肌梗死产率在不同条件下无显著差异。这些结果可用于相关实验室在某些情况下优化MI。
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来源期刊
CiteScore
5.00
自引率
11.50%
发文量
142
审稿时长
3 months
期刊介绍: The International Journal of Radiation Biology publishes original papers, reviews, current topic articles, technical notes/reports, and meeting reports on the effects of ionizing, UV and visible radiation, accelerated particles, electromagnetic fields, ultrasound, heat and related modalities. The focus is on the biological effects of such radiations: from radiation chemistry to the spectrum of responses of living organisms and underlying mechanisms, including genetic abnormalities, repair phenomena, cell death, dose modifying agents and tissue responses. Application of basic studies to medical uses of radiation extends the coverage to practical problems such as physical and chemical adjuvants which improve the effectiveness of radiation in cancer therapy. Assessment of the hazards of low doses of radiation is also considered.
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